肺腺癌细胞系耐药及凋亡相关基因的表达和逆转

目的 探讨顺铂耐药肺腺癌细胞系Ａ５４９＾ＤＤＰ的耐药，凋亡相关基因的表达及其与亲代细胞Ａ５４９的差异，观察差异表达基因的反义寡核苷酸的逆转作用。方法 将人工合成的反义，正义硫代脱氧寡核苷酸（Ｓ－ｏｌｉｇｏｄｌｅｏｘｙｎｕｃｌｅｏｔｉｄｅ，Ｓ－ＯＤＮ）经脂质体包后转染Ａ５４９＾ＤＤＰ细胞，应用ＲＴ－ＰＣＲ检测耐药及凋亡相关基因ｍＲＮＡ水平，免疫细胞化学及流式细胞－抗体检测相应蛋白及增殖抗原Ｋｉ－６７

Expression and reversion of drug resistance and apoptosis related genes of a DDP resistant lung adenocarcionma cell line A_(549)~(DDP)

OBJECTIVE: To study the expression of drug resistance- and apoptosis-related genes of A549DDP cells as compared to the parental cell line A549, and its reversion by antisense s-oligodeoxynucleotide (S-ODN) of the differentially expressed genes. METHODS: Sense and antisense S-ODN were transferred into A549DDP cells by lipofectin. Expression of genes related to drug resistance and apoptosis was examined by RT-PCR, immunocytochemistry and flow cytometry. Apoptosis was identified by DNA electrophoresis and TUNEL, and cell growth by MTT uptake. RESULTS: The expression of bcl-2 was positive and that of MRP at mRNA and protein levels was increased in A549DDP cells compared to A549 cells. MDR1, c-myc and TOPO II were similarly expressed in the two cell lines. Both cell lines were negative for c-erbB-2 expression. In A549DDP cells, the expression of bcl-2 and MRP was significantly inhibited by respective antisense S-ODN. Antisense S-ODN could also significantly inhibit proliferation of A549DDP cells, and promote cell apoptosis by reducing its resistance to cisplatin. CONCLUSION: Bcl-2 and MRP genes are responsible for the induced resistance of A549DDP cells to cisplatin.